Transgenic Mouse Models Shared Resource, Oregon Health and Science University, 3181 SW Sam Jackson Park Road, Portland, OR, 97239, USA.
In Vitro Cell Dev Biol Anim. 2024 Mar;60(3):300-306. doi: 10.1007/s11626-024-00884-5. Epub 2024 Mar 20.
The culture of preimplantation embryos in vitro is an important method for human and mouse reproductive technology. This study aims to investigate the influence of different conditions of culture media on the preimplantation stage of mouse embryos cultured in vitro, and monitor the post-implantation development of new mice after embryo transfer to surrogate females. We demonstrated here that mouse embryos cultured in vitro in fresh M16, KSOM, Global, and HTF embryo culture media from one cell to the blastocyst stage and the subsequent embryo transfer to surrogate females are able to proceed through post-implantation development and, after birth, develop into healthy mice. However, culture of embryos in differently aged media shows various (often unpredictable) results. To find the optimal storage conditions of culture media, we suggest that the freezing and long-term storage of these media at - 80°C will not influence the quality of the media. To test this hypothesis, we grew embryos from one cell to blastocysts in vitro in the selected media after thawing and subsequently transferring them to surrogate females. Embryo culture in these four media after thawing does not affect preimplantation and postnatal mouse development. Thus, we have shown that storage of embryo culture media at low temperature (- 80°C) does not impact the quality of the media, and subsequently, it can be used for the culture of embryos for the full preimplantation period, the same as in fresh media.
体外培养胚胎的文化是人类和老鼠生殖技术的重要方法。本研究旨在探讨不同的培养条件对体外培养的老鼠胚胎着床前阶段的影响，并监测胚胎转移到代孕雌性后的新老鼠的着床后发育情况。我们在这里证明，在新鲜的 M16、KSOM、Global 和 HTF 胚胎培养液中，老鼠胚胎从单细胞培养到囊胚阶段，以及随后的胚胎转移到代孕雌性，可以进行着床后发育，并在出生后发育成健康的老鼠。然而，胚胎在不同老化程度的培养液中的培养显示出各种（通常是不可预测的）结果。为了找到培养条件的最佳储存条件，我们建议将这些培养液在-80°C 下冷冻和长期储存不会影响培养液的质量。为了验证这一假设，我们在解冻后将胚胎从单细胞培养到囊胚，然后将其转移到代孕雌性中。在解冻后，这四种培养液中的胚胎培养不会影响老鼠的着床前和出生后发育。因此，我们已经表明，胚胎培养液的低温（-80°C）储存不会影响培养液的质量，随后可以用于培养胚胎的整个着床前阶段，就像在新鲜培养液中一样。
